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Stratagene ExSite Site DirectedMutagenesis Kit
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Posted 04 May 2001 - 09:00 PM
I'm having a problem with Stratagene's ExSite Site directed mutagenesiskit. I'm trying to delete about 500bp from a plasmid. The deletion worksOK, but in all clones, I get a single extra nucleotide inserted betweenthe two oligo sequences (i.e. in place of the delted sequence). This is usuallyan A ot T, but afew have been Gs. Stratagene suggest that the Pfu polishingstep may have failed, but I have tried several Pfus with the same result. It also happens with 3different primer pairs. Has anyone seen this before, and how did you getround it?